Amplification of a cytochrome P450 gene is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae.
Amplification of a cytochrome P450 gene is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae.
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DOI：
10.1371/journal.pgen.1000999
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发表时间：
2010-06-24
期刊：
影响因子：
4.5
通讯作者：
Bass C
中科院分区：
文献类型：
--
作者：
Puinean AM;Foster SP;Oliphant L;Denholm I;Field LM;Millar NS;Williamson MS;Bass C
蚜虫Myzus persicae是一种全球性的重要作物害虫，其已经进化出对几乎所有类别的杀虫剂的高水平抗性。迄今为止，烟碱类杀虫剂，一类经济上重要的杀虫剂，靶向烟碱乙酰胆碱受体（nAChRs），仍然是一种有效的控制措施，然而，最近的报告，在抗性M。桃蚜对这类化学品的长期功效构成威胁。本研究采用生物学、生物化学和基因组学方法，对烟草对烟碱类杀虫剂的抗性机制进行了研究。对一株抗性M.桃克隆（5191 A）的研究表明，P450介导的解毒作用在抗性中起主要作用，尽管其他机制也可能起作用。微阵列分析，使用与M中所有已知解毒基因相对应的探针填充的阵列。结果表明，P450基因（CYP 6CY 3）在桃蚜中的组成型过表达（22倍），定量PCR结果表明，P450基因的过表达至少部分是由于基因扩增所致。这是第一次报告的P450基因扩增事件与杀虫剂抗性在农业上重要的昆虫害虫。微阵列分析还显示了编码表皮蛋白的几种基因序列的过表达（2-16倍），人工喂养试验和使用放射性标记杀虫剂的体内渗透试验提供了减少表皮渗透在类烟碱抗性中的作用的直接证据。相反，受体放射性配体结合研究和nAChR亚基基因的核苷酸测序表明，靶位点的变化是不太可能有助于抗性的烟碱类杀虫剂在M。桃蚜。 M.在世界许多地区，由于其大的宿主范围以及其通过直接进食和通过传播植物病毒引起的损害，桃是经济上最重要的蚜虫害虫。该种已对大多数杀虫剂产生了抗药性;尽管迄今为止，类烟碱仍是一种有效的防治措施，但最近关于M.桃蚜对这类化学品的长期效力构成威胁。对其他昆虫物种的研究表明，抗药性可以通过增加代谢酶（如细胞色素P450）的产量来产生，这些酶可以使杀虫剂解毒。在这项研究中，我们利用基因组学的最新进展来量化所有编码解毒酶的基因在耐药菌株中的表达。桃蚜。我们发现抗性与单个P450基因的过度表达相关，我们表明这是由于基因扩增。我们还表明，角质层蛋白的过度表达和杀虫剂通过角质层的渗透减少也可能在抗性中发挥作用。这些发现将影响旨在减缓或防止耐药性发展的管理战略的设计和监测。
The aphid Myzus persicae is a globally significant crop pest that has evolved high levels of resistance to almost all classes of insecticide. To date, the neonicotinoids, an economically important class of insecticides that target nicotinic acetylcholine receptors (nAChRs), have remained an effective control measure; however, recent reports of resistance in M. persicae represent a threat to the long-term efficacy of this chemical class. In this study, the mechanisms underlying resistance to the neonicotinoid insecticides were investigated using biological, biochemical, and genomic approaches. Bioassays on a resistant M. persicae clone (5191A) suggested that P450-mediated detoxification plays a primary role in resistance, although additional mechanism(s) may also contribute. Microarray analysis, using an array populated with probes corresponding to all known detoxification genes in M. persicae, revealed constitutive over-expression (22-fold) of a single P450 gene (CYP6CY3); and quantitative PCR showed that the over-expression is due, at least in part, to gene amplification. This is the first report of a P450 gene amplification event associated with insecticide resistance in an agriculturally important insect pest. The microarray analysis also showed over-expression of several gene sequences that encode cuticular proteins (2–16-fold), and artificial feeding assays and in vivo penetration assays using radiolabeled insecticide provided direct evidence of a role for reduced cuticular penetration in neonicotinoid resistance. Conversely, receptor radioligand binding studies and nucleotide sequencing of nAChR subunit genes suggest that target-site changes are unlikely to contribute to resistance to neonicotinoid insecticides in M. persicae. M. persicae is the most economically important aphid pest in many regions of the world due to its large host range and the damage it causes through direct feeding and through the transmission of plant viruses. This species has developed resistance to most classes of insecticide; and although, to date, the neonicotinoids have remained an effective control measure, recent reports of resistance in M. persicae represent a threat to the long-term effectiveness of this chemical class. Work on other insect species has shown that resistance can arise through increased production of metabolic enzymes (such as cytochrome P450s) that detoxify the insecticide. In this study we made use of recent advances in genomics to quantify the expression of all genes encoding detoxification enzymes in a resistant strain of M. persicae. We found resistance correlated with the over-expression of a single P450 gene, and we show that this is due to gene amplification. We also showed that over-expression of cuticular proteins and reduced penetration of insecticide through the cuticle may also play a role in resistance. These findings will influence the design and monitoring of management strategies that aim to slow or prevent the development of resistance.
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